TIM-3/Galectin-9 interaction and glutamine metabolism in AML cell lines, HL-60 and THP-1
Abstract Background T cell immunoglobulin and mucin-domain containing-3 (TIM-3) is a cell surface molecule that was first discovered on T cells. However, recent studies revealed that it is also highly expressed in acute myeloid leukemia (AML) cells and it is related to AML progression. As, Glutamine...
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BMC
2024-01-01
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| Series: | BMC Cancer |
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| Online Access: | https://doi.org/10.1186/s12885-024-11898-3 |
| _version_ | 1797276480114262016 |
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| author | Hooriyeh Shapourian Mustafa Ghanadian Nahid Eskandari Abolfazl Shokouhi Gülderen Yanikkaya Demirel Alexandr V. Bazhin Mazdak Ganjalikhani-Hakemi |
| author_facet | Hooriyeh Shapourian Mustafa Ghanadian Nahid Eskandari Abolfazl Shokouhi Gülderen Yanikkaya Demirel Alexandr V. Bazhin Mazdak Ganjalikhani-Hakemi |
| author_sort | Hooriyeh Shapourian |
| collection | DOAJ |
| description | Abstract Background T cell immunoglobulin and mucin-domain containing-3 (TIM-3) is a cell surface molecule that was first discovered on T cells. However, recent studies revealed that it is also highly expressed in acute myeloid leukemia (AML) cells and it is related to AML progression. As, Glutamine appears to play a prominent role in malignant tumor progression, especially in their myeloid group, therefore, in this study we aimed to evaluate the relation between TIM-3/Galectin-9 axis and glutamine metabolism in two types of AML cell lines, HL-60 and THP-1. Methods Cell lines were cultured in RPMI 1640 which supplemented with 10% FBS and 1% antibiotics. 24, 48, and 72 h after addition of recombinant Galectin-9 (Gal-9), RT-qPCR analysis, RP-HPLC and gas chromatography techniques were performed to evaluate the expression of glutaminase (GLS), glutamate dehydrogenase (GDH) enzymes, concentration of metabolites; Glutamate (Glu) and alpha-ketoglutarate (α-KG) in glutaminolysis pathway, respectively. Western blotting and MTT assay were used to detect expression of mammalian target of rapamycin complex (mTORC) as signaling factor, GLS protein and cell proliferation rate, respectively. Results The most mRNA expression of GLS and GDH in HL-60 cells was seen at 72 h after Gal-9 treatment (p = 0.001, p = 0.0001) and in THP-1 cell line was observed at 24 h after Gal-9 addition (p = 0.001, p = 0.0001). The most mTORC and GLS protein expression in HL-60 and THP-1 cells was observed at 72 and 24 h after Gal-9 treatment (p = 0.0001), respectively. MTT assay revealed that Gal-9 could promote cell proliferation rate in both cell lines (p = 0.001). Glu concentration in HL-60 and α-KG concentration in both HL-60 (p = 0.03) and THP-1 (p = 0.0001) cell lines had a decreasing trend. But, Glu concentration had an increasing trend in THP-1 cell line (p = 0.0001). Conclusion Taken together, this study suggests TIM-3/Gal-9 interaction could promote glutamine metabolism in HL-60 and THP-1 cells and resulting in AML development. |
| first_indexed | 2024-03-07T15:28:47Z |
| format | Article |
| id | doaj.art-784e5866227444c7bc445fc4f7feff9f |
| institution | Directory Open Access Journal |
| issn | 1471-2407 |
| language | English |
| last_indexed | 2024-03-07T15:28:47Z |
| publishDate | 2024-01-01 |
| publisher | BMC |
| record_format | Article |
| series | BMC Cancer |
| spelling | doaj.art-784e5866227444c7bc445fc4f7feff9f2024-03-05T16:32:43ZengBMCBMC Cancer1471-24072024-01-0124111510.1186/s12885-024-11898-3TIM-3/Galectin-9 interaction and glutamine metabolism in AML cell lines, HL-60 and THP-1Hooriyeh Shapourian0Mustafa Ghanadian1Nahid Eskandari2Abolfazl Shokouhi3Gülderen Yanikkaya Demirel4Alexandr V. Bazhin5Mazdak Ganjalikhani-Hakemi6Department of Immunology, Faculty of Medicine, Isfahan University of Medical SciencesDepartment of Pharmacognosy, School of Pharmacy and Pharmaceutical Sciences, Isfahan University of Medical SciencesDepartment of Immunology, Faculty of Medicine, Isfahan University of Medical SciencesDepartment of Endocrine and metabolism research center, Isfahan University of Medical SciencesDepartment of Immunology, Faculty of Medicine, Yeditepe UniversityDepartment of General, Visceral and Transplant Surgery, Ludwig Maximilians University of MunichDepartment of Immunology, Faculty of Medicine, Isfahan University of Medical SciencesAbstract Background T cell immunoglobulin and mucin-domain containing-3 (TIM-3) is a cell surface molecule that was first discovered on T cells. However, recent studies revealed that it is also highly expressed in acute myeloid leukemia (AML) cells and it is related to AML progression. As, Glutamine appears to play a prominent role in malignant tumor progression, especially in their myeloid group, therefore, in this study we aimed to evaluate the relation between TIM-3/Galectin-9 axis and glutamine metabolism in two types of AML cell lines, HL-60 and THP-1. Methods Cell lines were cultured in RPMI 1640 which supplemented with 10% FBS and 1% antibiotics. 24, 48, and 72 h after addition of recombinant Galectin-9 (Gal-9), RT-qPCR analysis, RP-HPLC and gas chromatography techniques were performed to evaluate the expression of glutaminase (GLS), glutamate dehydrogenase (GDH) enzymes, concentration of metabolites; Glutamate (Glu) and alpha-ketoglutarate (α-KG) in glutaminolysis pathway, respectively. Western blotting and MTT assay were used to detect expression of mammalian target of rapamycin complex (mTORC) as signaling factor, GLS protein and cell proliferation rate, respectively. Results The most mRNA expression of GLS and GDH in HL-60 cells was seen at 72 h after Gal-9 treatment (p = 0.001, p = 0.0001) and in THP-1 cell line was observed at 24 h after Gal-9 addition (p = 0.001, p = 0.0001). The most mTORC and GLS protein expression in HL-60 and THP-1 cells was observed at 72 and 24 h after Gal-9 treatment (p = 0.0001), respectively. MTT assay revealed that Gal-9 could promote cell proliferation rate in both cell lines (p = 0.001). Glu concentration in HL-60 and α-KG concentration in both HL-60 (p = 0.03) and THP-1 (p = 0.0001) cell lines had a decreasing trend. But, Glu concentration had an increasing trend in THP-1 cell line (p = 0.0001). Conclusion Taken together, this study suggests TIM-3/Gal-9 interaction could promote glutamine metabolism in HL-60 and THP-1 cells and resulting in AML development.https://doi.org/10.1186/s12885-024-11898-3Acute myeloid leukemia (AML)T cell immunoglobulin and mucin-domain containing-3 (TIM-3)Galectin-9 (Gal-9)Glutamine metabolismGlutaminase (GLS)Glutamate dehydrogenase (GDH) |
| spellingShingle | Hooriyeh Shapourian Mustafa Ghanadian Nahid Eskandari Abolfazl Shokouhi Gülderen Yanikkaya Demirel Alexandr V. Bazhin Mazdak Ganjalikhani-Hakemi TIM-3/Galectin-9 interaction and glutamine metabolism in AML cell lines, HL-60 and THP-1 BMC Cancer Acute myeloid leukemia (AML) T cell immunoglobulin and mucin-domain containing-3 (TIM-3) Galectin-9 (Gal-9) Glutamine metabolism Glutaminase (GLS) Glutamate dehydrogenase (GDH) |
| title | TIM-3/Galectin-9 interaction and glutamine metabolism in AML cell lines, HL-60 and THP-1 |
| title_full | TIM-3/Galectin-9 interaction and glutamine metabolism in AML cell lines, HL-60 and THP-1 |
| title_fullStr | TIM-3/Galectin-9 interaction and glutamine metabolism in AML cell lines, HL-60 and THP-1 |
| title_full_unstemmed | TIM-3/Galectin-9 interaction and glutamine metabolism in AML cell lines, HL-60 and THP-1 |
| title_short | TIM-3/Galectin-9 interaction and glutamine metabolism in AML cell lines, HL-60 and THP-1 |
| title_sort | tim 3 galectin 9 interaction and glutamine metabolism in aml cell lines hl 60 and thp 1 |
| topic | Acute myeloid leukemia (AML) T cell immunoglobulin and mucin-domain containing-3 (TIM-3) Galectin-9 (Gal-9) Glutamine metabolism Glutaminase (GLS) Glutamate dehydrogenase (GDH) |
| url | https://doi.org/10.1186/s12885-024-11898-3 |
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